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A technology of Mycoplasma hyopneumoniae and detection kit, which can be applied to measurement devices, instruments, scientific instruments, etc., can solve the problems of influence of detection results, low expression level of target protein, and difficulty in large-scale popularization and use.
Active Publication Date: 2008-08-06
CHINA INST OF VETERINARY DRUG CONTROL
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[0004] For the detection of swine panting disease, there is currently only an indirect hemagglutination detection reagent for Mycoplasma hyopneumoniae in China. However, due to the high manufacturing cost, the detection results are easily affected by factors such as operators, environment and materials, and it is difficult to popularize it on a large scale.
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[0046] Prepared according to the following formula: sodiumchloride 8.0g, KH 2 PO 4 0.3g, Na 2 HPO 4 .12H 2 O 5.33g, KCl 0.2g, add water for injection to 900mL, add 5%-10% skimmed milkpowder and fully dissolve, adjust pH to 7.0-7.2, add 0.01%-0.05% sodiumazide, add water for injection to volume 1000mL.
[0047] (2) Preparation of enzyme-binding working solution
[0048] The enzyme-binding working solution is a commercially available horseradish peroxidase-labeled rabbit anti-pig antibody (referred to as the enzyme-labeled secondary antibody). The verification test is carried out according to the recommended dilution factor in the instructions for use, and the correction is made according to the test results. If the recommended dilution factor is 20000×, use working concentrations of 2500×, 5000×, 10000×, 20000× and 40000× to add to the secondary antibody diluent, and perform ELISA assays resp...
[0072] Preparation before the experiment: Dilute the 10× washing solution with distilled water to make 1× washing working solution. Dilute the sample to be tested 50 times with diluent.
[0074] (2) Add each sample to be tested into the test sample well, 100 μL per well, put it in a self-sealing bag and place it at room temperature for 30 minutes;
[0075] (3) Pour off the liquid in each well and wash with 1× washing solution, 350 μL per well, and wash 3 to 5 times;
[0076] (4) Add enzyme-labeled secondary antibody working solution, 100 μL per well, pack it in a self-sealing bag, place it at room temperature for 30 minutes, and repeat step 3);
[0077] (5) Add 10 μL of color-developing working solution, and develop color at room temperature for 15 minu...
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technical field [0001] The invention relates to a kit for detecting animal infectious diseases, which belongs to the field of veterinary biological products. Background technique [0002] Mycoplasma hypneumoniae (Mycoplasma hypneumoniae, Mhp) is a pathogenic microorganism that causes swine mycoplasma pneumonia (also known as panting disease or porcine enzootic pneumonia). The disease is characterized by contact, highly contagious, chronic, high morbidity and low mortality. It is mainly manifested as cough and dyspnea. Anatomical findings are mainly lung lesions, especially the heart lobe, middle lobe and apex of the two lungs. Pancreatoid and sarcoid features appear in the leaves. Its main harm is that it causes the growth of pigs to be hindered and the feed conversion rate to drop sharply. In addition, its high morbidity rate seriously affects the economic benefits of raising pigs. Studies in recent years have found that mixed infection of Mycoplasma hyopneumoniae with po...
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